The study of optimal conditions of electroporation in Gluconacetobacter xylinum

Abstract

The effect of electroporation conditions of Gluconacetobacter xylinum CGMCC 2955 was systematically investigated in this paper. The enzymolysis condition for removing bacterial cellulose of Gluconacetobacter xylinum was optimized by Response Surface Method, the result showed as follows: cellulase concentration 1046.4 IU/mL, enzymolysis duration 93.8 min at 30.6°C. Optimum conditions including growth stage of the strain, electroshock voltage, concentration and preservation of competent cell were defined for the electroporation of CGMCC 2955 with plasmid pDN19LacΩ. The result was showed that the highest transformation efficiency was up to 8.76×10 5 CFU/μg DNA under the optimium conditions in which the competent cells were collected at 2d and concentration to about 1×10 8 CFU/mL, the mixture of the competent cells and plasmid pDN19LacΩ was eletroporated at 2.4kV. The transformation efficiency is significantly reduced on account of -70°C preservation of competent cell. © 2012 Springer-Verlag GmbH.